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Real-time quantitative PCR analysis showed that low phosphate (Pi) stress significantly induced the expression of <i>ZmPHR1</i> and <i>ZmPHR2</i> in maize seedling stage, and candidate gene association analysis further revealed the close association of these two genes with root traits under Pi stress conditions. Transgenic plants overexpressing <i>ZmPHR1</i> and <i>ZmPHR2</i> in <i>Arabidopsis</i> show a significant increase in lateral root number, fresh weight and total phosphorus accumulation under low-Pi stress. Besides, CHIP-PCR experiments identified target genes involved in hormone regulation, metal ion transport and homeostasis, phosphatase encoding, and photosynthesis, providing new insights into the biological functions of <i>ZmPHR1</i> and <i>ZmPHR2</i>. Furthermore, our study showed that ZmPHR1 interacts with six SPX domain-only proteins (ZmSPXs) in maize, while ZmPHR2 interacts with five of these proteins. <i>ZmPHR1</i> and <i>ZmPHR2</i> expression was repressed in low Pi conditions, but was up-regulated in <i>ZmSPX1</i> knockout material, according to our study of transgenic seedlings overexpressing <i>ZmSPX1</i> in maize. We identified downstream target genes involved in the phosphorus signaling pathway, which are mainly involved in plant-pathogen interactions, ascorbic acid and arabinose metabolism, and ABC transporter proteins, by RNA-seq analysis of transgenic seedlings grown under low Pi stress for 7 days. Collectively, these results provide important clues to elucidate the role and functional significance of <i>ZmPHR1</i> and <i>ZmPHR2</i> under low Pi stress and also provide insights into understand the molecular mechanism of phosphorus homeostasis in maize.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s11032-024-01508-2.</p>","PeriodicalId":18769,"journal":{"name":"Molecular Breeding","volume":null,"pages":null},"PeriodicalIF":2.6000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11442720/pdf/","citationCount":"0","resultStr":"{\"title\":\"Functional analysis of <i>ZmPHR1</i> and <i>ZmPHR2</i> under low-phosphate stress in maize.\",\"authors\":\"Hongmei Hu, Yikai Wang, Haixu Zhong, Binyang Li, Jingxiao Qi, Yarong Wang, Jin Liu, Shuhao Zhang, Haiying Zhang, Bowen Luo, Xiao Zhang, Zhi Nie, Hongkai Zhang, Duojiang Gao, Shiqiang Gao, Dan Liu, Ling Wu, Shibin Gao\",\"doi\":\"10.1007/s11032-024-01508-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The PHOSPHATE STARVATION RESPONSE REGULATOR (PHR) plays a crucial regulatory role in plants during the process of responding to phosphate starvation. 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引用次数: 0
摘要
磷酸盐饥饿响应调节因子(PHR)在植物响应磷酸盐饥饿的过程中起着至关重要的调节作用。本研究结合反向遗传学和生物技术,研究了玉米幼苗中ZmPHR1和ZmPHR2的功能,包括含有Myb_DNA_banding和Myb_CC-LHEQLE结构域的蛋白质。系统进化分析表明,ZmPHR1和ZmPHR2与AtPHR1和OsPHR2具有高度同源性,并具有核定位和转录自激活的特征。实时定量 PCR 分析表明,低磷酸盐(Pi)胁迫显著诱导玉米苗期 ZmPHR1 和 ZmPHR2 的表达,候选基因关联分析进一步揭示了这两个基因与 Pi 胁迫条件下根系性状的密切关联。在拟南芥中过表达 ZmPHR1 和 ZmPHR2 的转基因植株在低 Pi 胁迫下的侧根数量、鲜重和总磷积累都有显著增加。此外,CHIP-PCR 实验还发现了涉及激素调控、金属离子转运和稳态、磷酸酶编码和光合作用的靶基因,为了解 ZmPHR1 和 ZmPHR2 的生物学功能提供了新的视角。此外,我们的研究还发现,ZmPHR1 与玉米中的六个纯 SPX 结构域蛋白(ZmSPXs)相互作用,而 ZmPHR2 与其中的五个蛋白相互作用。根据我们对玉米过表达 ZmSPX1 的转基因幼苗的研究,ZmPHR1 和 ZmPHR2 的表达在低 Pi 条件下受到抑制,但在 ZmSPX1 基因敲除的材料中则上调。通过对在低 Pi 胁迫下生长 7 天的转基因幼苗进行 RNA-seq 分析,我们确定了磷信号通路的下游靶基因,这些基因主要涉及植物与病原体的相互作用、抗坏血酸和阿拉伯糖代谢以及 ABC 转运体蛋白。总之,这些结果为阐明 ZmPHR1 和 ZmPHR2 在低 Pi 胁迫下的作用和功能意义提供了重要线索,也为了解玉米磷平衡的分子机制提供了见解:在线版本包含补充材料,见 10.1007/s11032-024-01508-2。
Functional analysis of ZmPHR1 and ZmPHR2 under low-phosphate stress in maize.
The PHOSPHATE STARVATION RESPONSE REGULATOR (PHR) plays a crucial regulatory role in plants during the process of responding to phosphate starvation. In this study, we combined reverse genetics and biotechnology to investigate the function of ZmPHR1 and ZmPHR2, including proteins containing the Myb_DNA_banding and Myb_CC-LHEQLE structural domains, in maize seedlings. Phylogenetic analysis revealed that ZmPHR1 and ZmPHR2 have high homology with AtPHR1 and OsPHR2, and share the characteristic features of nuclear localisation and transcriptional self-activation. Real-time quantitative PCR analysis showed that low phosphate (Pi) stress significantly induced the expression of ZmPHR1 and ZmPHR2 in maize seedling stage, and candidate gene association analysis further revealed the close association of these two genes with root traits under Pi stress conditions. Transgenic plants overexpressing ZmPHR1 and ZmPHR2 in Arabidopsis show a significant increase in lateral root number, fresh weight and total phosphorus accumulation under low-Pi stress. Besides, CHIP-PCR experiments identified target genes involved in hormone regulation, metal ion transport and homeostasis, phosphatase encoding, and photosynthesis, providing new insights into the biological functions of ZmPHR1 and ZmPHR2. Furthermore, our study showed that ZmPHR1 interacts with six SPX domain-only proteins (ZmSPXs) in maize, while ZmPHR2 interacts with five of these proteins. ZmPHR1 and ZmPHR2 expression was repressed in low Pi conditions, but was up-regulated in ZmSPX1 knockout material, according to our study of transgenic seedlings overexpressing ZmSPX1 in maize. We identified downstream target genes involved in the phosphorus signaling pathway, which are mainly involved in plant-pathogen interactions, ascorbic acid and arabinose metabolism, and ABC transporter proteins, by RNA-seq analysis of transgenic seedlings grown under low Pi stress for 7 days. Collectively, these results provide important clues to elucidate the role and functional significance of ZmPHR1 and ZmPHR2 under low Pi stress and also provide insights into understand the molecular mechanism of phosphorus homeostasis in maize.
Supplementary information: The online version contains supplementary material available at 10.1007/s11032-024-01508-2.
期刊介绍:
Molecular Breeding is an international journal publishing papers on applications of plant molecular biology, i.e., research most likely leading to practical applications. The practical applications might relate to the Developing as well as the industrialised World and have demonstrable benefits for the seed industry, farmers, processing industry, the environment and the consumer.
All papers published should contribute to the understanding and progress of modern plant breeding, encompassing the scientific disciplines of molecular biology, biochemistry, genetics, physiology, pathology, plant breeding, and ecology among others.
Molecular Breeding welcomes the following categories of papers: full papers, short communications, papers describing novel methods and review papers. All submission will be subject to peer review ensuring the highest possible scientific quality standards.
Molecular Breeding core areas:
Molecular Breeding will consider manuscripts describing contemporary methods of molecular genetics and genomic analysis, structural and functional genomics in crops, proteomics and metabolic profiling, abiotic stress and field evaluation of transgenic crops containing particular traits. Manuscripts on marker assisted breeding are also of major interest, in particular novel approaches and new results of marker assisted breeding, QTL cloning, integration of conventional and marker assisted breeding, and QTL studies in crop plants.